Fermented Barley Extracts with Lactobacillus plantarum dy-1 Rich in Vanillic Acid Modulate Glucose Consumption in Human HepG2 Cells

Objective To investigate the effect of fermented barley extracts with Lactobacillus plantarum dy-1（LFBE） for modulating glucose consumption in HepG2 cells via miR-212 regulation. Methods Hepatocellular carcinoma（HepG2） cells were treated with palmitate. After 12 h, palmitate-induced HepG2 cells were treated with LFBE and its main components. Changes in glucose consumption, proinflammatory cytokine secretion, and miRNA-212 expression in HepG2 cells was observed. Results Treatment with LFBE rich in vanillic acid（VA） increased glucose consumption and reduced proinflammatory cytokine secretion in HepG2 cells. LFBE and VA normalized the upregulation of miR-212, which led to the upregulation of dual-specificity phosphatase-9（DUSP9）, a direct target of miR-212, at both protein and mR NA levels. Downregulation of miR-212 markedly increased glucose consumption and reduced proinflammatory cytokine secretion by enhancing DUSP9 expression. Conclusion The results showed the benefit of LFBE and miR-212 downregulation in modulating glucose consumption and reducing proinflammatory cytokine secretion by targeting DUSP9. VA in LFBE was a strong regulator of palmitate-induced abnormal glucose consumption in HepG2 cells and can be a primary mediator.

Comparison of a glucose consumption based method with the CLSI M38-A method for testing antifungal susceptibility of Trichophyton rubrum and Trichophyton mentagrophytes

Background The prevalence of dermatophytoses and the development of new antifungal agents has focused interest on susceptibility tests of dermatophytes. The method used universally for susceptibility tests of dermatophytes was published as document （M38-A） in 2002 by the Clinical and Laboratory Standards Institute （CLSI）, dealing with the standardization of susceptibility tests in filamentous fungi, though not including dermatophytes especially. However, it is not a very practical method for the clinical laboratory in routine susceptibility testing. In this test, we developed a novel rapid susceptibility assay --glucose consumption method （GCM） for dermatophytes. Methods In this study, we investigated the antifungal susceptibilities of dermatophytes to itraconazole （ITC）, voriconazole （VOC）, econazole nitrate （ECN） and terbinafine （TBF） by glucose consumption method （GCM）, in comparison to the Clinical and Laboratory Standards Institute （CLSI） M38-A method. Twenty-eight dermatophyte isolates, including Trichophyton rubrum （T. rubrum） （n=-14） and Trichophyton mentagrophytes （T. mentagrophytes） （n=-14）, were tested. In the GCM, the minimum inhibitory concentrations （MICs） were determined spectrophotometrically at 490 nm after addition of enzyme substrate color mix. For the CLSI method, the MICs were determined visually. Results Comparison revealed best agreement for TBF against T. mentagrophytes and T. rubrum, since MIC range, MIC50, and MIC90 were identical from two methods. However, for ITC and VOC, GCM showed wider MIC ranges and higher MICs than CLSI methods in most isolates. For ECN against T. rubrum, high MICs were tested by GCM （0.125-16 pg/ml） but not M38-A method （0.5-1 IJg/ml）. The overall agreements for all isolates between the two methods within one dilution and two dilutions for ITC, VOC, ECN and TBF was 53.6% and 75.0%, 57.1% and 75.0%, 82.1% and 89.3%, and 85.7 and 85.7%, respectively. Conclusion Measurement of glucose uptake can predict the susceptibility of T. rubrum and T. mentagrophytes to ECN and TBF.

Syringaresinol-4-O-β-D-glucoside alters lipid and glucose metabolism in HepG2 cells and C2C12 myotubes

Syringaresinol-4-O-β-D-glucoside(SSG), a furofuran-type lignan, was found to modulate lipid and glucose metabolism through an activity screen of lipid accumulation and glucose consumption, and was therefore considered as a promising candidate for the prevention and treatment of metabolic disorder,especially in lipid and glucose metabolic homeostasis. In this study, the effects of SSG on lipogenesis and glucose consumption in Hep G2 cells and C2C12 myotubes were further investigated. Treatment with SSG significantly inhibited lipid accumulation by oil red O staining and reduced the intracellular contents of total lipid, cholesterol and triglyceride in Hep G2 cells. No effect was observed on cell viability in the MTT assay at concentrations of 0.1–10 μmol/L. SSG also increased glucose consumption by Hep G2 cells and glucose uptake by C2C12 myotubes. Furthermore, real-time quantitative PCR revealed that the beneficial effects were associated with the down-regulation of sterol regulatory element-binding proteins-1c,-2(SREBP-1c,-2), fatty acid synthase(FAS), acetyl CoA carboxylase(ACC) and hydroxyl methylglutaryl CoA reductase(HMGR), and up-regulation of peroxisome proliferator-activated receptors alpha and gamma(PPARα and PPARγ). SSG also significantly elevated transcription activity of PPARγtested by luciferase assay. These results suggest that SSG is an effective regulator of lipogenesis and glucose consumption and might be a candidate for further research in the prevention and treatment of lipid and glucose metabolic diseases.

Chlorella sp.-ameliorated undesirable microenvironment promotes diabetic wound healing

Chronic diabetic wound remains a critical challenge suffering from the complicated negative microenvironments,such as high-glucose,excessive reactive oxygen species(ROS),hypoxia and malnutrition.Unfortunately,few strategies have been developed to ameliorate the multiple microenvironments simultaneously.In this study,Chlorella sp.(Chlorella)hydrogels were prepared against diabetic wounds.In vitro experiments demonstrated that living Chlorella could produce dissolved oxygen by photosynthesis,actively consume glucose and deplete ROS with the inherent antioxidants,during the daytime.At night,Chlorella was inactivated in situ by chlorine dioxide with human-body harmless concentration to utilize its abundant contents.It was verified in vitro that the inactivated-Chlorella could supply nutrition,relieve inflammation and terminate the oxygen-consumption of Chlorella-respiration.The advantages of living Chlorella and its contents were integrated ingeniously.The abovementioned functions were proven to accelerate cell proliferation,migration and angiogenesis in vitro.Then,streptozotocininduced diabetic mice were employed for further validation.The in vivo outcomes confirmed that Chlorella could ameliorate the undesirable microenvironments,including hypoxia,high-glucose,excessiveROS and chronic inflammation,thereby synergistically promoting tissue regeneration.Given the results above,Chlorella is considered as a tailor-made therapeutic strategy for diabetic wound healing.

AstragalosideⅣameliorates insulin induced insulin resistance in HepG2 cells through reactive oxygen species mediated c-Jun N-terminal kinase pathway

OBJECTIVE:To investigate the effects and elucidate the mechanism of Astragaloside IV(AS-IV)for insulin resistance(IR)and type 2 diabet es mellitus(T2DM).METHODS:CCK8 kit was used to detect cell viability,glucose detection kit was used to detect the concentration of glucose in cell supernatant,reactive oxygen species(ROS)detection kit and Western blot were used to explore the mechanism of Astragaloside IV(AS-IV)in improving IR.A diabetic rat model was also established by feeding high sugar and fat diet and streptozotocin(STZ)injection.After treatment with AS-IV,rosiglitazone(ROZ),or normal saline,the fasting blood glucose(FBG),C peptide(C-P),tumor necrosis factor-α(TNF-α),interleukin-6(IL-6)and the glucose tolerance were assessed.RESULTS:AS-IV could effectively reduce the content of ROS and increase the glucose uptake in high insulintreated IR-type HepG 2 cells.The results of molecular mechanisms indicated that AS-IV could improve insulin resistance by reducing JNK phosphorylation and regulating c-Jun N-terminal kinase(JNK)downstream protein expression.Additionally,AS-IV could significantly reduce the levels of FBG,TNF-α,IL-6 and the glucose tolerance in diabetic rats(P<0.05 or<0.01).The high and medium dose groups of AS-IV could significantly increase the C-P levels in diabetic rats(P<0.05 or<0.01).CONCLUSIONS:Our results indicated that AS-IV improve liver IR through the JNK pathway and ROS,which meant a new molecular target for the treatment of diabetes.The AS-IV also helped to prevent and improved the insulin resistance of rats.

2,3-Seco and 3-nor guaianolides from Achillea alpina with antidiabetic activity

In this study,we presented the isolation and characterization of eight novel seco-guaianolide sesquiterpenoids(1-8)and two known guaianolide derivatives(9 and 10),from the aerial part of Achillea alpina L..Compounds 1-3 were identified as guaianolides bearing an oxygen insertion at the 2,3 position,while compounds 4-8 belonged to a group of special 3-nor guaianolide sesquiterpenoids.The structural elucidation of 1-8,including their absolute configurations,were accomplished by a combination of spectroscopic data analysis and quantum electronic circular dichroism(ECD)calculations.To evaluate the potential antidiabetic activity of compounds 1-10,we investigated their effects on glucose consumption in palmitic acid(PA)-mediated HepG2-insulin resistance(IR)cells.Among the tested compounds,compound 7 demonstrated the most pronounced ability to reverse IR.Moreover,a mechanistic investigation revealed that compound 7 exerted its antidiabetic effect by reducing the production of the pro-inflammatory cytokine IL-1β,which was achieved through the suppression of the NLRP3 pathway.

Discovery and development of tricyclic matrinic derivatives as anti-diabetic candidates by AMPKαactivation

We found compound 12N-p-trifluoromethylbenzenesulfonyl matrinane(1)was a potent anti-diabetic agent.Thirty-five tricyclic matrinic derivatives were synthesized and determined for their stimulatory effects on glucose consumption in L6 myotubes,taking 1 as the lead.In high-fat diet(HFD)and STZ induced diabetic mice,9a significantly lowers blood glucose,improves glucose tolerance,and especially alleviates diabetic nephropathy and islet damage.Mechanism study indicates that 9a simultaneously targets mitochondrial complex I to increase AMP/ATP ratio,as well as liver kinase B1(LKB1)and calcium/calmodulindependent protein kinase(Ca MKK),which synergistically activates AMPKαand then stimulates glucose transporter 4(GLUT4)membrane translocation and 2-deoxyglucose(2-DG)uptake to exert anti-diabetic efficacy.Therefore,compound 9a with a novel structure is a promising anti-diabetic candidate with the advantage of multiple-target mechanism,worthy of further investigation.